人CXCL8 knockout PC-3 cell line
Human CXCL8 knockout PC-3 cell line
- Advanced Validation
- 了解详情
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CXCL8 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 22% 11 bp deletion, 24% 7 bp deletion, 54% 2 bp deletion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
查看别名
(Ala-IL-8)77, (Ser-IL-8)72, 9.00E+03, Beta thromboglobulin like protein, C-X-C motif chemokine 8, CEF-4, CXCL8, Emoctakin, GCP-1, GCP/IL-8 protein I, GCP/IL-8 protein II, GCP/IL-8 protein III, GCP/IL-8 protein IV, GCP/IL-8 protein V, GCP/IL-8 protein VI, Granulocyte chemotactic protein 1, IL-8(1-77), IL-8(9-77), IL8/NAP1 form I, IL8/NAP1 form II, IL8/NAP1 form III, IL8/NAP1 form IV, IL8/NAP1 form V, IL8/NAP1 form VI, IL8_HUMAN, Inteleukin 8, LECT, LUCT, LYNAP, Lymphocyte-derived neutrophil-activating factor, MDNCF, MDNCF-b, MDNCF-c, MONAP, Monocyte-derived neutrophil chemotactic factor, Monocyte-derived neutrophil-activating peptide, NAP-1, Neutrophil activating peptide 1, Neutrophil-activating factor, Neutrophil-activating protein 1, Protein 3-10C, SCYB 8, Small inducible cytokine subfamily B member 8, T-cell chemotactic factor, TSG 1, chemokine, CXC motif, ligand 8
- WB
Lab
Western blot - Human CXCL8 knockout PC-3 cell line (AB273743)
Lanes 1 - 6 : Merged signal (red and green). Green - ab235584 observed at 10 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
ab235584 was shown to react with IL-8 in wild-type PC-3 cells in Western blot with loss of signal observed in CXCL8 knockout cell line ab273743 (knockout cell lysate ab275520). Wild-type PC-3 and CXCL8 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab235584 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 ° at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-IL-8 antibody [EPR22994-255] (<a href='/products/primary-antibodies/il-8-antibody-epr22994-255-ab235584'>ab235584</a>) at 1/1000 dilution
Lane 1:
Wild-type PC-3 Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5 μg/ml, 5 h) cell lysate at 30 µg
Lane 2:
Wild-type PC-3 LPS-treated (2 μg/ml, 6 h) with Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>) (5 μg/ml, 5 h) cell lysate at 30 µg
Lane 2:
Western blot - Human CXCL8 knockout PC-3 cell line (ab273743)
Lane 3:
CXCL8 knockout PC-3 Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5 μg/ml, 5 h) cell lysate at 30 µg
Lane 4:
CXCL8 knockout PC-3 LPS-treated (2 μg/ml, 6 h) with Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>) (5 μg/ml, 5 h) cell lysate at 30 µg
Lane 5:
A431 cell lysate at 30 µg
Lane 6:
HCT116 cell lysate at 30 µg
Predicted band size: 11 kDa
Observed band size: 10 kDa
false
- sELISA
Lab
Sandwich ELISA - Human CXCL8 knockout PC-3 cell line (AB273743)
Human IL-8 concentration was interpolated from the standard curve. Supernatants from cell culture samples were serially diluted and assessed by the Human IL-8 ELISA kit (ab214030). Wild-type PC-3 cells and CXCL8 knockout PC-3 cells (ab273743) were assessed in duplicate (n=2) and were either treated with 2 μg/ml LPS for 6 hours to induce expression of IL-8 or not treated with LPS. Data are represented as the mean and error bars represent standard deviation.
- NGS
Supplier Data
Next Generation Sequencing - Human CXCL8 knockout PC-3 cell line (AB273743)
Allele-2 : 7bp deletion in exon 2.
- NGS
Supplier Data
Next Generation Sequencing - Human CXCL8 knockout PC-3 cell line (AB273743)
Allele-3 : 2bp deletion in exon 2.
- NGS
Supplier Data
Next Generation Sequencing - Human CXCL8 knockout PC-3 cell line (AB273743)
Allele-1 : 11bp deletion in exon 2.
- ICC/IF
Supplier Data
Immunocytochemistry/ Immunofluorescence - Human CXCL8 knockout PC-3 cell line (AB273743)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized CXCL8 KO PC-3 (CXCL8 knockout human prostate adenocarcinoma epithelial cell), ab273743 cells labelling IL-8 with ab322732 at 1/2000 (0.246 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/ml dilution (Green).
Confocal image showing increased cytoplasmic staining in wildype PC-3 cells treated with Lipopolysaccharide (2 ug/ml) and Brefeldin A (5 ug/ml) for 5 hours (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2ug/ml dilution.
反应性数据
产品详情
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
规格
性能和储存信息
基因名称
基因编辑类型
基因编辑方法
敲除验证
运输条件
推荐的短期储存条件
推荐的长期储存条件
处理步骤
初始处理指南
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
传代培养指南
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
培养基
F-12K + 10% FBS
低温储藏试剂
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
补充信息
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
IL-8 functions as a chemoattractant for neutrophils and lymphocytes facilitating their movement towards the site of infection or injury. It does not form part of a larger protein complex but operates individually to enhance immune cell migration. IL-8 possesses unique binding motifs allowing it to interact with specific receptors namely CXCR1 and CXCR2 on target cells. This binding triggers cellular responses leading to effective immune surveillance and response to inflammatory stimuli.
Pathways
IL-8 operates within important inflammatory and immune response pathways. It forms a part of the NF-κB signaling cascade which activates in response to stress signals promoting the expression of other inflammatory mediators. Additionally it engages in the mitogen-activated protein kinase (MAPK) pathway influencing cellular responses such as proliferation and differentiation. The interaction of IL-8 with these pathways highlights its role in modulating immune responses and highlights its interaction with other proteins such as TNF-α and IL-1β.
细胞培养
生物安全等级
EU: 1 US: 1
贴壁/悬浮
Adherent
性别
Male
搭配使用产品
Primary Antibodies
AB242515
Anti-IL-8 antibody [EPR19358-37] - BSA and Azide free (Capture)
BOND RX™ Validated|RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IL-8 antibody [EPR19358-37] - BSA and Azide free (Capture) (AB242515)](https://content.abcam.com/products/images/il-8-antibody-epr19358-37-bsa-and-azide-free-capture-ab242515--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img441719.jpg)
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